Download Purified Mouse Anti-Human Retinoblastoma

BD Pharmingen™
Technical Data Sheet
Purified Mouse Anti-Human Retinoblastoma Protein
Product Information
Material Number:
Alternate Name:
Size:
Concentration:
Clone:
Immunogen:
Isotype:
Reactivity:
Target MW:
Storage Buffer:
554144
RB
0.1 mg
0.5 mg/ml
XZ55
Recombinant carboxy-terminal human RB protein aa. 387-928
Mouse IgG1
QC Testing: Human
Reported: Mouse, Chicken, Xenopus, Quail
105-116 kDa
Aqueous buffered solution containing ≤0.09% sodium azide.
Description
The retinoblastoma gene encodes a nuclear phosphoprotein (Rb or p110[Rb]) which is expressed in most normal cells of vertebrates and acts
as a tumor suppressor gene product. An underphosphorylated form of RB is mainly found in resting or fully differentiated cells, whereas the
hyperphosphorylated form is present in proliferating cells. Only the underphosphorylated form of RB binds specifically to viral oncogenes
such as SV40 large T, adenviral EIA and HPV-E7. This interaction may partially contribute to the transforming activity of these viral
oncoproteins. Rb also interacts with several cyclins including A, D, and E as well as the transcriptional activator E2F. The importance of these
interactions for the biological function of Rb is still being elucidated.
XZ55 recognizes an epitope located between amino acids 443-622 of human Rb. It cross-reacts with chicken, mouse and Xenopus Rb, and a
putative quail Rb.A recombinant carboxy-terminal (amino acids 387-928) human Rb protein was used as immunogen.
Western blot analysis of Rb in MOLT-4 human leukemia
cell lysate. The XZ55 antibody (Cat. No. 554144)
recognizes phosphorylated and underphosphorylated forms
of Rb (~110-116 kDa).
Preparation and Storage
The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography.
Store undiluted at 4°C.
Application Notes
Application
Western blot
Routinely Tested
Immunoprecipitation
Reported
Gel shift
Reported
Recommended Assay Procedure:
Application includes western blot analysis (1-2 µg/ml). Rb migrates as multiple closely-spaced bands between approximately 110-116 kDa when
sized on denaturing polyacrylamide gels (i.e. by SDS-PAGE). The different bands represent different Rb phosphorylation states, the higher
molecular weight bands are more highly phosphorylated than the lower molecular weight bands. The level of phosphorylation is cell cycle
554144 Rev. 1
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dependent, and may also be cell type dependent (not all forms are seen in all cell types that express Rb). Gel conditions influence the actual
number of bands observed. In cases where optimal band separation is desired, use a 7 to 10% non-gradient long (≥ 12 inches) gel. MOLT-4
human leukemia cells (ATCC CRL-1582) are suggested as a positive control. Additional applications not routinely tested at BD Biosciences
Pharmingen include immunoprecipitation (1-2 µg/ml) and electrophoretic mobility gel shift assays (EMSA).
Suggested Companion Products
Catalog Number
554002
Name
HRP Goat Anti-Mouse Ig
Size
1.0 ml
Clone
(none)
Product Notices
1.
2.
3.
Since applications vary, each investigator should titrate the reagent to obtain optimal results.
Please refer to www.bdbiosciences.com/pharmingen/protocols for technical protocols.
Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before
discarding to avoid accumulation of potentially explosive deposits in plumbing.
References
Guilhot C, Benchaibi M, Flechon JE, Samarut J. The 12S adenoviral E1A protein immortalizes avian cells and interacts with the avian RB product. Oncogene.
1993; 8(3):619-624.(Clone-specific: Western blot)
Hollingsworth RE Jr, Chen PL, Lee WH. Integration of cell cycle control with transcriptional regulation by the retinoblastoma protein. Curr Opin Cell Biol
Curr Opin Cell Biol. 1993; 5(2):194-200.(Biology)
Hu QJ, Bautista C, Edwards GM, Defeo-Jones D, Jones RE, Harlow E. Antibodies specific for the human retinoblastoma protein identify a family of related
polypeptides. Mol Cell Biol. 1991; 11(11):5792-5799.(Clone-specific: Immunoprecipitation, Western blot)
Livingston DM. Functional analysis of the retinoblastoma gene product and of RB-SV40 T antigen complexes. Cancer Surv. 1992; 12:153-160.(Biology)
Riley DJ, Lee EY, Lee WH. The retinoblastoma protein: more than a tumor suppressor. Annu Rev Cell Biol. 1994; 10:1-29.(Biology)
Shirodkar S, Ewen M, DeCaprio JA, Morgan J, Livingston DM, Chittenden T. The transcription factor E2F interacts with the retinoblastoma product and a
p107-cyclin A complex in a cell cycle-regulated manner. Cell. 1992; 68(1):157-166.(Clone-specific: Gel shift)
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